Examining the impact of laboratory housing temperature on murine CD28 and the response to anti-PD-1

检查实验室外壳温度对小鼠 CD28 的影响以及抗 PD-1 的反应

• 批准号: 10751424
• 负责人: Caitlin M James
• 金额: $ 3.25万
• 依托单位: ROSWELL PARK CANCER INSTITUTE CORP
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-09-01 至 2025-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10751424
• 关键词: Address; Affect; Animal Housing; Animal Model; Animals; Basal metabolic rate; Biomedical Research; CD28 gene; CD8-Positive T-Lymphocytes; CD8B1 gene; Cancer Model; Caring; Chronic; Clinic; Clinical; Data; Development; Down-Regulation; Flow Cytometry; Genetically Engineered Mouse; Goals; Guidelines; Harvest; House mice; Housing; Human; Immune; Immune checkpoint inhibitor; Immunologics; Immunotherapy; Impairment; In Vitro; Institution; Knockout Mice; Knowledge; Laboratories; Laboratory Animal Science; Laboratory Animals; Malignant Neoplasms; Measures; Mediating; Medical; Modeling; Molecular; Molecular Target; Mus; Nerve; Norepinephrine; Outcome; Pathway interactions; Patients; Phenotype; Physiological; Play; Pre-Clinical Model; Process; Publishing; Receptor Signaling; Reproducibility; Research; Research Proposals; Rodent; Role; Signal Induction; Signal Transduction; Standardization; Surface; Sympathetic Nervous System; T-Cell Activation; T-Lymphocyte; Temperature; Testing; Thermogenesis; Training; Tumor Immunity; Tumor-Infiltrating Lymphocytes; Work; adrenergic stress; anti-PD-1; anti-tumor immune response; anticancer research; beta-adrenergic receptor; cancer therapy; career; cold stress; design; effector T cell; environmental stressor; experience; falls; functional disability; guide for the care and use of laboratory animals; human disease; human model; improved; in vivo; insight; mRNA Expression; melanoma; mouse model; novel therapeutics; pre-clinical; preclinical study; protein expression; receptor; response; success; therapy development; translational potential; treatment response; tumor; tumor growth; tumor immunology

Project Abstract/Summary For decades, animals in biomedical research have yielded significant scientific and medical breakthroughs by generating the essential preclinical data that ultimately support the discovery and development of treatments for human diseases, including cancer. However, while we to rely on animal models to investigate the complexity of cancer and cancer therapies, these preclinical studies have alarmingly low success in reproducibility, and even lower preclinical-to-clinical success rates. As per the Guide for the Care and Use of Laboratory Animals 8th Edition, research institutions have standardized, minimum guidelines for the housing, husbandry, and overall care for laboratory animals that they must adhere to. A mildly cool ambient temperature is a critical aspect of animal housing that has been shown to elicit significant physiological changes to research rodents, driven by the activation of the sympathetic nervous system and increased β- adrenergic receptor (β-AR) signaling as a result of the systemic release of norepinephrine. This is due to the compensatory response, known as non-shivering thermogenesis, employed by rodents housed at temperatures that fall below their thermoneutral zone (which is the range of ambient temperatures at which heat generated by basal metabolism is sufficient for maintaining homeostatic core temperature. ) Our lab has previously established that standard (ST), subthermoneutral laboratory housing temperatures result in significant impairment to the murine CD8+ T cell-dependent anti-tumor immune response compared to mice house at thermoneutral temperatures (TT). Additionally, we have shown that the immune checkpoint inhibitor αPD-1, an immunotherapy that has recently seen success as a front-line approach to treating cancers like melanoma, has improved efficacy in treating tumor-bearing mice housed at TT in a β-AR-dependent manor. Although published and preliminary data indicate a role for the co-receptor, CD28, in the diminished anti-tumor function of CD8+ T cells as a result of increased β-AR signaling, a gap exists in our understanding of the mechanisms underlying the reduced CD8+ T cell activation and effector function in mice housed at ST. Therefore, we propose using genetically engineered mouse models to precisely interrogate CD28 signaling and test hypothesis that standard housing temperatures impairs CD8+ T cell anti-tumor immunity and the in vivo efficacy of αPD-1 via impaired CD28 co-stimulation. We will use in vitro and in vivo approaches to examine the effects of housing temperature on CD8+ T cell CD28 expression and signaling, as well as tumor- infiltrating lymphocytes in mice treated with αPD-1 therapy. The studies outline in this proposal have the potential to identify a previously undefined mechanism by which subthermoneutral laboratory animal housing temperatures influence experimental outcomes of cancer and immunotherapy models, while also characterizing a widely underappreciated variable that exists in our animal models.

项目摘要/摘要 几十年来，生物医学研究中的动物通过 生成最终支持治疗发现和开发的基本临床前数据 对于包括癌症在内的人类疾病。但是，尽管我们依靠动物模型来研究 这些临床前研究的复杂性和癌症疗法的复杂性令人震惊 可重复性，甚至较低的临床前的成功率。根据护理和使用指南 实验室动物第8版，研究机构已经标准化了住房的最低指南， 畜牧业，以及必须遵守的实验动物的整体护理。温和的环境 温度是动物外壳的关键方面，已证明会引起大量的生理 研究啮齿动物的变化是由交感神经系统的激活驱动的，并增加了β- 肾上腺素的全身释放导致肾上腺素受体（β-AR）信号传导。这是由于 补偿性响应，称为非动荡的热发生，由啮齿动物所用 温度低于其热区域（这是环境温度的范围 基本代谢产生的热量足以维持稳态核心温度。 ）我们的实验室有 以前确定标准（ST），亚热实验室住房温度导致 与小鼠相比 嗜热温度（TT）的房屋。此外，我们已经表明了免疫抑制剂 αPD-1，一种免疫疗法，最近将成功作为治疗癌症的前线方法 黑色素瘤在治疗β-AR依赖性庄园中含有TT的肿瘤小鼠方面的效率提高了。 尽管发表和初步数据表明了抗肿瘤中共受体CD28的作用 CD8+ T细胞的功能由于β-AR信号的增加而导致的，我们对 降低CD8+ T细胞激活和效应子功能的机制。 因此，我们建议使用一般设计的鼠标模型精确询问CD28信号传导 并检验假设，标准壳体温度会损害CD8+ T细胞抗肿瘤免疫组织切和IN 通过受损的CD28共刺激，αPD-1的体内效率。我们将使用体外和体内方法 检查住房温度对CD8+ T细胞CD28表达和信号传导的影响，以及肿瘤 用αPD-1治疗治疗的小鼠浸润淋巴细胞。该提案中的研究大纲具有 潜力识别以前未定义的机制 温度会影响癌症和免疫疗法模型的实验结果，同时也 表征我们动物模型中存在的广泛被低估的变量。